Plant derived seed extract rich in essential fatty acids derived from perilla seed: composition of matter, manufacturing process and use

ABSTRACT

A composition of matter comprises a shelf stable, super critical, CO 2  fluid extracted seed oil derived from a cracked biomass of  perilla frutescens , the seed oil comprising from about 60 to about 95 percent w/w of PUFAs in a ratio of from about 4:1 to about 6:1 alpha-linolenic acid (ALA) to linoleic acid (LA) and a mixture of selected antioxidants.

RELATED APPLICATION(S)

This is a continuation-in-part application of application Ser. No.12/419,321 filed Apr. 7, 2009, which is based upon provisionalapplication Ser. No. 61/043,773 filed Apr. 10, 2008, the disclosureswhich are hereby incorporated by reference in their entirety.

FIELD OF THE INVENTION

This invention relates to plant-derived seed extracts and methods ofmaking same.

BACKGROUND OF THE INVENTION

It is well known in the literature that Polyunsaturated Fatty Acids(PUFAs) of all types are highly susceptible to peroxide, free radicaland light induced degradation including rancification and polymerizationmaking them unsuitable for human consumption. For example, it is wellknown that flax seed oil, also known as linseed oil, readily undergoesfree radical oxidation to advantageously form polymeric surfacesincluding oil based paints, hard furniture finishes and linoleumflooring. In addition, many companies offer flax seed oil for humanconsumption as a dietary supplement or food ingredient because of thehigh levels of PUFAs found in raw flax seed and its expeller pressedoils and more particularly Alpha-Linolenic Acid (ALA) and Linolenic Acid(LA). Many flax seed oil product labels suggest that the product must berefrigerated at all times due to the instability of such PUFAs in flaxseed oil. Careful examination of the majority of commercially availableflax seed oils obtained by expeller pressing, including those typicallystored under refrigerated conditions, unfortunately reveals that theyare unfit for human use based on their measured Peroxide Values (PVs).Such PV values above 3 meq/Kg (milliequivalents/gram) are deemed notsuitable for salad oil applications and PV values above 10 meq/Kg aredeemed to be unsuitable for human use because the measured PV value maybe an early indicator of rancidity and free radical induced degradation.On the other hand, PV values taken alone do not adequately characterizesuch oils since a low PV value can also be associated with PUFA's thathave already gone through the rancification process. Typical testing hasrevealed flax seed oil products sold for human consumption with observedPV's as high as 130 meq/kg also characterized with the odor associatedwith short chain aldehydes that make such oils “rancid” to olefactorysenses.

Most raw seed based oils in common cooking and baking use, such assoybean, corn and canola seed oils naturally contain enough PUFAs makingthem unsuitable, without further processing, for use as cooking oils.Therefore unless such PUFA containing raw seed oils are hydrogenated tofully saturated triglycerides using hydrogen and a catalyst prior totheir use in cooking applications, they are considered to be unfit foruse as cooking oils. These oils are typically first isolated by, forexample, expeller pressing the appropriate seed. The crude seed oil isthen filtered to remove biomass. The resulting oil, containingsignificant levels of PUFAs, is then catalytically hydrogenated toreduce the PUFA content to levels suitable for use of the resulting oilin cooking applications. If the hydrogenation process is incomplete,however, the resulting mixtures are found to contain both undesirableheat labile PUFAs that quickly undergo rancification to small chainaldehydes in the resulting heated cooking oil as well as unsaturatedtrans-fatty acids which are believed to be detrimental to animal andespecially human health.

Therefore, those skilled in the art will recognize the great difficultyin producing a shelf stable PUFA mixture wherein the PUFA content is ashigh as 70% wt/wt of the resulting seed extract from a natural seedsource that then exhibits extraordinary room temperature stability.

Commonly assigned and copending patent application Ser. No. 12/419,321discloses a Salvia hispanica L. derived seed oil extract composition ofmatter containing from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 ofALA to LA, 4-10% of C-18 mono-unsaturated fatty acid, 1-5% of C-18saturated fatty acid and 4-8% of C-16 saturated fatty acid in a mixedtriglyceride form stable at room temperature of 12-24 months containinga mixture of selected antioxidants. This chia derived composition has avery favorable ratio of ALA to LA (omega 3 to omega 6) of about 3.3:1 inone example. It is desirable to provide a composition of matter as aseed oil extract derived from other than chia and a method ofmanufacturing the seed oil that has equivalent or better PUFA contentand ratios of ALA to LA and is shelf stable and exhibits roomtemperature stability.

SUMMARY OF THE INVENTION

A perilla seed oil extract provides a healthy source of omega-3 andcontains a very favourable ratio of ALA to LA (omega-3 to omega-6) ofabout 6:1 in one example. The perilla oil is obtained by pressing theperilla seeds to initiate seed cracking, processing the cracked biomassusing supercritical CO² fluid extraction, and collecting the extract todeliver more of the higher molecular weight compounds.

A composition of matter comprises a shelf stable, supercritical, CO²fluid extracted seed oil derived from a cracked biomass of perillafrutescens, the seed oil comprising from about 60 to about 95 percentw/w of PUFAs in a ratio of from about 4:1 to about 6:1 alpha-linolenicacid (ALA) to linoleic acid (LA) and a mixture of selected antioxidants.

In one example the selected antioxidants comprise a synergistic mixtureof selected lipophilic and hydrophilic antioxidants. In another examplethe composition comprises lipophilic antioxidants either alone or incombination with at least one of: a) phenolic antioxidants including atleast one of sage, oregano, and rosemary; b) tocopherol, c)tocotrienol(s), d) carotenoids including at least one of astaxanthin,lutein, and zeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g)Butylated hydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i)Tertiary Butyl hydroquinone (TBHQ). In another example a hydrophilicantioxidant or sequesterant is added comprising hydrophilic phenolicantioxidants including at least one of grape seed extract, tea extracts,ascorbic acid, citric acid, tartaric acid, and malic acid.

In another example the peroxide value of the seed oil is under 10.0meq/Km. The composition can have supplemented to it docosahexaenoic acid(DHA) and/or eicosapentaenoic acid (EPA) in pectin or gelatin basedconfectionary dietary supplement delivery systems. The composition mayinclude EPA, DHA, docosapentaenoic acid (DPA) or gamma-linlolenic acid(GLA), fish oil, krill oil, krill oil concentrate, borage oil, eveningprimrose oil, olive oil or other plant, animal or algal based seed orfruit oils admixed therein.

The composition of matter in yet another example comprises from about 85to about 95 percent w/w of PUFAs. In another example, the compositionhas at least greater than 50 percent alpha-linolenic acid (ALA). Theseed oil is shelf stable at room temperature up to 32 months. In anotherexample the seed oil is derived from a premilled or flake-rolled crackedbiomass of perilla frutescens.

A method is also disclosed of mitigating or preventing cardiovasculardisease, arthritic pain and inflammation, platelet aggregation, ortreating dry eye syndrome, premenstrual symptoms or modifying immuneresponse in humans or animals by applying an effective amount of adietary supplement, food or beverage that has added thereto acomposition mixed therewith and comprising a supercritical, CO² fluidextracted seed oil derived from a cracked biomass of perilla frutescens,the seed oil comprising from about 60 to about 95 percent w/w of PUFAsin a ratio of from about 4:1 to about 6:1 alpha-linolenic acid (ALA) tolinoleic acid (LA) and a mixture of selected antioxidants.

In one example, an emulsifying agent is added. Nano- and/ormicro-particles of rice or sugarcane based policosanol can be added forproviding a heart healthy dietary supplement. Also, a stabilized oil canbe dispersed in a fruit juice concentrate, fruit puree or other waterbased flavoring in the presence of maltodextrin, or other carbohydrates,and an emulsifying or emulsion stabilization agent that is vacuum spraydried to form an amorphous or crystalline solid suitable for use as aflavoring ingredient carrying stabilized PUFAs in flavored dietarysupplements, foods and beverages.

Oil based flavors and fragrances suitable for use as an ingredient infoods, beverages and cosmetics can be added. Docosahexaenoic acid (DHA)and/or eicosapentaenoic acid (EPA) can be added to supplement, includingadding EPA, DHA, docosapentaenoic acid (DPA) or gamma-linlolenic acid(GLA), fish oil, krill oil, krill oil concentrate, borage oil, eveningprimrose oil, olive oil or other plant, animal or algal based seed orfruit oils.

A method of manufacturing a perilla frutescens derived seed oilcomprising from about 60 to about 95 percent w/w of PUFAs in a ratio offrom about 4:1 to about 6:1 alpha-linolenic acid (ALA) to linoleic acid(LA) is disclosed and comprises processing perilla frutescens seedpreferably in the absence of oxygen to obtain a biomass having a desiredparticle size distribution, subjecting the resulting biomass tosupercritical fluid CO² extraction, collecting a resulting seed oilfraction, and separating water in the fraction.

The method also comprises in one example treating any resulting seed oilfraction with antioxidants to impart a desired room temperaturestability. The method further comprises controlling the extent of oilextraction by particle size distribution. The method further comprisesadding propane in admixture with supercritical CO² in the supercriticalstate as an extraction solvent. In one example, the method furthercomprises extracting solvent using hexane extraction at or nearatmospheric pressures and the resulting boiling point of hexane in theabsence of oxygen, separating the resulting water from the oil/hexanemixture and removing the hexane solvent by distillation at or belowatmospheric pressure in the absence of oxygen.

The method further comprises adding lipophilic antioxidants to increasethe room temperature stability of the resulting oil. The method furthercomprises adding lipophilic antioxidants either alone or in combinationwith at least one of: a) phenolic antioxidants including at least one ofsage, oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d)carotenoids including at least one of astaxanthin, lutein, andzeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g) Butylatedhydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i) TertiaryButyl hydroquinone (TBHQ). The method further comprises treating anyresulting dewatered seed oil with bleaching clay or activated carbon.

The method further comprises adding a hydrophilic antioxidant orsequesterant comprising hydrophilic phenolic antioxidants including atleast one of grape seed extract, tea extracts, ascorbic acid, citricacid, tartaric acid, and malic acid. The method further comprisespremilling or roller press flaking the perilla frutescens seed to obtainthe desired particle size. The method further comprises treating thepremilled or roller press flaked seed with a lipophilic or hydrophilicantioxidant prior to solvent extraction. The method further comprisesprocessing the perilla frutescens seed with or without the addition ofhydrophilic or lipophilic antioxidants during the particle sizingprocess. The perilla frutescens derived seed oil is stable at roomtemperature for about 12 to about 32 months. The method furthercomprises performing supercritical extraction in the presence oflipophilic and or hydrophilic antioxidants.

BRIEF DESCRIPTION OF THE DRAWINGS

Other objects, features and advantages of the present invention willbecome apparent from the detailed description of the invention whichfollows, when considered in light of the accompanying drawings in which:

FIG. 1 is a flowchart illustrating a production diagram as a flowchartfor producing omega-3 chia seed oil such as sold under the tradenameChia Gold™ by Valensa International of Eustis, Fla.

FIG. 2 is another production diagram as a flowchart showing basic stepsfor manufacturing a perilla seed extract in accordance with anon-limiting example.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention will now be described more fully hereinafter. Thisinvention may, however, be embodied in many different forms and shouldnot be construed as limited to the embodiments set forth herein. Rather,these embodiments are provided so that this disclosure will be thoroughand complete, and will fully convey the scope of the invention to thoseskilled in the art.

Commonly assigned and copending parent application Ser. No. 12/419,321discloses a room temperature, shelf stable mixture of an approximate3.1:1 to about 3.3:1 mixture of alpha-linolenic acid (“ALA”, “Omega-3polyunsaturated fatty acid” (“PUFA”)) to linoleic acid (“LA”, “Omega-6PUFA) that has been prepared in the presence of limited amounts ofsaturated and mono-unsaturated fatty acids as their mixed triglyceridesby the use of either supercritical fluid CO² solvent extraction ofpremilled Salvia hispanica L. seed alone, and more particularly,supercritical fluid CO² solvent extraction in the presence of mixturesof hydrophilic and lipophilic antioxidants, or, by the use of a commonorganic solvent extraction such as hexane or by the use of expellerpressing techniques. The supercritical CO² extraction is preferred.

Such Omega-3 and Omega-6 PUFAs are well known as essential fatty acidsin man and many animals, which are useful in humans and animals inpromoting, for example, a heart healthy condition in man. It is alsowell known, however, that PUFAs are extremely susceptible to rapid,uncontrollable free radical mediated degradation.

The composition of matter disclosed in the copending '321 applicationincludes a supercritical CO² Salvia hispanica L. derived seed oilcomprising from 60-88% PUFAs in a ratio of from about 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of C-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and comprising a mixture ofselected antioxidants.

The composition of matter includes dietary supplement ingredients suchas docosahexaenoic acid (DHA) and/or eicosapentaenoic acid (EPA) inpectin or gelatin based confectionary dietary supplement deliverysystems. EPA, DHA, docosahexaenoic acid (DPA) or gamma-linlolenic acid(GLA), fish oil, krill oil, krill oil concentrate, borage oil, eveningprimrose oil, olive oil or other plant, animal or algal based seed orfruit oils are admixed therein either alone or in combination.Lipophilic antioxidants are added either alone or in combination with atleast one of a) phenolic antioxidants including at least one of sage,oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d) carotenoidsincluding at least one of astaxanthin, lutein, and zeaxanthin; e)ascorbylacetate; f) ascorbylpalmitate g) Butylated hydroxytoluene (BHT);h) Docosapentaenoic Acid (BHA) and i) Tertiary Butyl hydroquinone(TBHQ). As disclosed, a hydrophilic antioxidant or sequesterant includeshydrophilic phenolic antioxidants including at least one of grape seedextract, tea extracts, ascorbic acid, citric acid, tartaric acid, andmalic acid.

A method of manufacturing and method of using the composition is alsoset forth in the '321 application.

As further disclosed in this incorporated by reference '321 application,this PUFA rich seed oil extract is prepared from Salvia hispanica L.seed which contains one of nature's more favorable seed basedconcentrations and ratios for the essential fatty acids, and morespecifically, the essential fatty acids ALA and LA in a ratio ofapproximately 3.3:1 as a mixture of ALA and LA that is stable at roomtemperature for long periods of time when desirably and appropriatelytreated with antioxidants either before, during, or after (or anycombination thereof). A shelf life of 12-24 months has been found.

Such oils are used either alone or advantageously in combination withother ingredients, for example, algae, plant or fish derivedalpha-linolenic acid (ALA) or linoleic acid (LA) metabolites such aseicosapentaenoic acid (EPA), docosapentaenoic acid (DPA),gamma-linlolenic acid (GLA) or docosahexaenoic acid (DHA) or anycombination thereof, incorporated into appropriate foods, beverages ordietary supplements for the prevention or mitigation of such diseases ascardiovascular disease, arthritis, pain, blood clotting, dry eyes andbrain health.

Such disease mitigation has been associated with the competitive controlof the LA metabolic cascade and the resulting metabolic cascade productsfrom LA metabolism known commonly as eicosanoids, such as the series 2and 3 prostaglandins and thromboxanes, the series 4 leucotrienes andlipoxins and the series 5 leuotrienes all of which are potent plateletaggregators and/or inhibitors, pro-inflammatories, vasodilators,bronchoconstrictors, or anti-asthmatics and the like.

The consumption of ALA has been shown to be a very effective competitivesubstrate of delta-6 desaturase, which is known to be the rate limitingenzymatic step in both ALA and LA metabolism to the metabolic productsdiscussed above.

Attempted extraction of Saliva hispanica L. unmilled seed, usingsupercritical CO² even at extraordinarily high pressures of 1000 bar orhexane solvent at atmospheric pressures, yields very little, if any,seed oil, therefore the seed must be milled prior to extraction. Theextent of the milling, as measured by particle size distribution, isadvantageous to the extraction process in accordance with a non-limitingaspect since the higher the surface area, the higher will be theefficiency and completeness of the extraction process by either organicsolvent based ox supercritical fluid based processes. In addition, it isoften advantageous to mill the seed in a blanket of inert gas such asnitrogen to prevent per-oxidative processes from taking place that wouldotherwise be initiated in the presence of air or oxygen and light.

In one embodiment, Salvia Hispanica L. whole seed is either firstcommutated in a standard knife or hammer mill or more preferably rollermilled, preferably under a cold nitrogen atmosphere, to produce acracked seed biomass. The seed biomass is preferably treated with one ormore hydrophilic and/or lipophilic antioxidants by mixing theantioxidants to the resulting biomass. In another embodiment, theantioxidant may be advantageously added to the seed prior to or duringthe milling process or at the point of extraction without pre-blendingthe antioxidants evenly throughout the resulting biomass due to thenature of the extraction process. The biomass is then transferred to asupercritical fluid extraction unit for separation of the seed oil fromthe cracked or flake-rolled biomass.

Alternatively, the pre-prepared biomass can be transferred to a commonhexane solvent extractor, or an expeller press for example, and the oilextracted from the biomass accordingly. Preferably either process isconducted in the absence of oxygen or air.

The supercritical fluid extraction of the milled seed admixed withhydrophilic and/or lipophilic antioxidants is accomplished by subjectingthe premilled cracked or flake-rolled seed to supercritical CO², or CO²and propane as a co-solvent, or supercritical propane alone at from40-1000 bar at from 30-100 Deg. C. More preferably the seed oil isextracted from the biomass between 50-800 bar at 50-90 deg. C. in suchCO² amounts measured in kgs/kg of biomass and for such times as may berequired to extract large portions of the seed oil content from thebiomass. In addition, entrainment solvents can be added to thesupercritical fluid to further enhance the efficacy of such extractions.For example, supercritical carbon dioxide extraction of the biomass canbe enhanced by the addition of propane to the supercritical extractionfluid.

The resulting seed oil dissolved in supercritical solvent(s) is nextallowed to fractionate in two separate pressure step-down stagesallowing the collection of a light and heavy fraction of seed oilextract. This light fraction also contains water that has beenco-extracted from the seed mass. The resulting seed oil, afterdegassing, is separated from any water that may have been carried overduring the extraction of the biomass containing the water. The lightfraction of the seed oil extract is rich in taste and odor componentsand may be admixed with the heavy fraction or may be discarded dependingon the desired product characteristics.

After separation of the water remaining in each fraction, the fractionsare then held under nitrogen or other inert gas and additional amountsof lipophilic and/or hydrophilic antioxidants may then be added. Inaddition, the resulting fractions may also be treated with bleachingclay, carbon and such other processing aids as may be required to renderthe oil suitable for its intended use in humans and animals.

The PV of the resulting seed oil extract is typically under 2.0 meq/Km,while accelerated decomposition, using a Rancimat instrument, remarkablyindicates an extrapolated room temperature shelf life of from about 1-2years. When the same process is repeated without the use ofantioxidants, the resulting PV is surprisingly under 10.0 meq/Kg mostprobably due to the use of supercritical CO² resulting in minimalexposure of the oil to oxygen species. However, the resulting oilquickly begins to build peroxide value in the presence of air even whenstored at temperatures of 0 Degs. C. In addition, such unstabilizedoils, under accelerated rancimat testing exhibit very poor stability toheat and oxygen unlike the rancimat performance observed in stabliziedoils derived from the process described above.

The resulting supercritical fluid seed oil extract of the inventioncontains from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of ALA:LA,4-10% of C18 mono-unsaturated fatty acid, 1-5% of C-18 saturated fattyacid and 4-8% C-16 saturated fatty acid composition in a mixedtriglyceride form depending on the seed source employed.

On the other hand, if the process described above is conducted withoutthe use of hydrophilic and/or lipophilic antioxidants, the resultingseed oil extract exhibits an initial low PV but accelerated stabilitytesting using a Rancimat instrument indicates an extrapolated roomtemperature shelf stability of less than two months.

The stability of the resulting oil at room temperature that ismanufactured without the use of added antioxidants cannot be easilyexplained because of the available levels of the powerful naturalantioxidants found in Salvia hispanica L. whole seed whose activity canbe easily measured in Oxygen Radical Absorbance Capacity (ORAC) units.Salvia hispanica L. has a measured ORAC number of 3000 micromoles TEORAC units/gram of seed and is known to contain such antioxidants asmyricetin, quercetin, kaempferol, caffeic acid, and chlorogenic acid. Inaddition, it is well known that the Salvia hispanica L. whole seed,unlike many other seeds bearing PUFA containing oil, exhibits a shelflife of at least 5 years due to its structure and the naturallyoccurring antioxidants available in the seed matrix.

In addition, cold pressing of Salvia hispanica L. whole seed alsoproduces unstable seed oil without careful addition of appropriateantioxidants to the seed prior to the expeller pressing process.

In a non-limiting example the composition of matter is formed from asupercritical CO² derived Salvia hispanica L. derived seed oilcomprising from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of C-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and comprising a mixture ofselected antioxidants.

It includes docosahexaenoic acid (DHA) and/or eicosapentaenoic acid(EPA) in pectin or gelatin based confectionary dietary supplementdelivery systems and in another aspect EPA, DHA, docosapentaenoic acid(DPA) or gamma-linlolenic acid (GLA), fish oil, krill oil, krill oilconcentrate, borage oil, evening primrose oil, olive oil or other plant,animal or algal based seed or fruit oils are admixed therein.

Lipophilic antioxidants are added either alone or in combination with atleast one of a) phenolic antioxidants including at least one of sage,oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d) carotenoidsincluding at least one of astaxanthin, lutein, and zeaxanthin; e)ascorbylacetate; f) ascorbylpalmitate g) Butylated hydroxytoluene (BHT);h) Docosapentaenoic Acid (BHA) and i) Tertiary Butyl hydroquinone(TBHQ). The hydrophilic antioxidant or sequesterant includes hydrophilicphenolic antioxidants including at least one of grape seed extract, teaextracts, ascorbic acid, citric acid, tartaric acid, and malic acid inanother aspect.

A method of manufacturing a Salvia hispanica L. derived seed oil inanother non-limiting example is set forth. The seed oil comprises from60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of alpha-linolenic acid(ALA) to linoleic acid (LA), 4-10% of C-18 mono-unsaturated fatty acid,1-5% of C-18 saturated fatty acid and 4-8% of C-16 saturated fatty acidin a mixed triglyceride form that is stable at room temperature for12-24 months and includes antioxidants. The method includes milling orroller press flaking Salvia hispanica L. seed in the absence of oxygento obtain a desired particle size distribution with or without theaddition of hydrophilic or lipophilic antioxidants during the particlesizing process. The resulting biomass is subjected to supercriticalfluid CO² extraction in the presence of lipophilic and/or hydrophilicantioxidants. Any resulting seed oil fractions are collected. The wateris separated in each fraction.

Any resulting seed oil fractions can be treated with additionalantioxidants to afford a desired room temperature stability. The extentof oil extraction can be controlled by particle size distribution of themilled or flaked seed. Propane can be added in mixture withsupercritical CO² in the supercritical state as an extraction solvent.In yet another aspect solvent can be extracted using hexane extractionat or near atmospheric pressures and the resulting boiling point ofhexane in the absence of oxygen, separating the resulting water from theoil/hexane mixture and removing the hexane solvent by distillation at orbelow atmospheric pressure in the absence of oxygen.

Lipophilic antioxidants can be added to increase the room temperaturestability of the resulting oil. The lipophilic antioxidants can be addedeither alone or in combination with at least one of a) phenolicantioxidants including at least one of sage, oregano, and rosemary; b)tocopherol, c) tocotrienol(s), d) carotenoids including at least one ofastaxanthin, lutein, and zeaxanthin; e) ascorbylacetate; f)ascorbylpalmitate g) Butylated hydroxytoluene (BHT); h) DocosapentaenoicAcid (BHA) and i) Tertiary Butyl hydroquinone (TBHQ). The resultingdewatered seed oil can be treated with bleaching clay or activatedcarbon.

Premilled or roller press flaked seed can be treated with a lipophilicor hydrophilic antioxidant(s) prior to solvent extraction. Thehydrophilic antioxidant or sequesterant can be formed from hydrophilicphenolic antioxidants including at least one of grape seed extract, teaextracts, ascorbic acid, citric acid, tartaric acid, and malic acid.

A method of mitigating or preventing cardiovascular disease, arthriticpain and inflammation, platelet aggregation, or treating dry eyesyndrome, premenstrual symptoms or modifying immune response in humansor animals is set forth by applying an effective amount of a dietarysupplement, food or beverage to which has been a composition mixedtherewith and comprising a Salvia hispanica L. derived seed oilcomprising from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of 0-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and includes antioxidants.

In one aspect, an emulsifying agent is added. In another aspect, nano-and/or micro-particles of rice or sugarcane based polycosanol aredispersed for enhancing a heart healthy dietary supplement. A stabilizedoil in a fruit juice concentrate, fruit puree or other water basedflavoring is dispersed in the presence of maltodextrin, or othercarbohydrates, and a suitable emulsifying or emulsion stabilizationagent that is vacuum spray dried to form an amorphous or crystallinesolid suitable for use as a flavoring ingredient carrying stabilizedPUFAs in flavored dietary supplements, foods and beverages. In yetanother aspect, oil based flavors and fragrances suitable for use as aningredient in foods, beverages and cosmetics are added. ALA and LA arealso added as essential fatty acids.

It has also been found that the use of a perilla seed oil extractinstead of the disclosed chia seed oil is advantageous and contains avery favorable ratio of ALA to LA (omega-3 to omega-6) of as high asabout 6:1 in some examples as compared to chia seed oil that istypically about 3.3:1 ALA to LA. Perilla seed oil extract as obtained,in accordance with a no-limiting example, is thus an even healthiersource of omega-3 than chia seed oil in various non-limiting examples.

FIG. 1 is a flowchart showing a production diagram for a sequence ofsteps for producing omega-3 chia seed oil such as sold under thetradename Chia Gold™ by Valensa International of Eustis, Fla. The salviahispanica seed is provided (block 100) and seed cracking occurs (block102) to form a cracked biomass. Various techniques for seed cracking andforming the biomass can be used as described above. The supercriticalCO² extraction (block 104) uses ultra high pressure carbon dioxideextraction technology and supplied CO² (block 106) such as the DEEPEXTRACT® manufacturing process developed by Valensa International ofEustis, Fla. The defatted seed as a flour, for example, (block 108) isproduced. Separation of the different portions occurs such as byfractionating the seed oil extract (block 110) as described above. Theextract is collected (block 112). Dewatering and decantation occurs(block 114) and antioxidants are added (block 116) such as the OTB® PerOxidation blocker system from Valensa International. Quality controlanalysis occurs (block 118) and the final oil packaged (block 120).

FIG. 2 shows a second production diagram as a flowchart for producingthe perilla seed oil extract in accordance with a non-limiting example.The process begins with a source of the perilla seed also known asperilla frutescens seed (block 130). Similar to the process with theomega-3 chia seed oil, seed cracking occurs (block 132) to form acracked seed biomass followed by the extraction (block 134) using thesupercritical CO² extraction and supplying carbon dioxide (block 136) toproduce the defatted seed (block 138) that is partially or whollydefatted perilla seed as a cake residue with virtually no fat or oil.The other portion is the oil and the CO² is reclaimed (block 140).Similar to the omega-3 chia seed oil, the extract is collected (block142) and dewatering and decantation occurs (block 144). Antioxidantssuch as the OTB® components as described before are added (block 146)and the quality control analysis occurs (block 148) followed bypackaging (block 150).

The perilla seed extract can have a range of values for its fatty acidprofile. Total fatty acids, peroxide values and other component valuesare described in Table 1 below showing an analysis of perilla seedextract and various parameters, specifications and results as anon-limiting example. Results can vary of course for different samples.

TABLE 1 Ingredients: Perilla (Perilla frutescens (L) Britton) Seed Oil,O2B ™ Botanical peroxidation blocker including refined nonGMO soybeanmixed tocopherols and spice extract. Parameter Specification ResultDescription Clear yellow oil, pourable Conforms at room temperature OdorMild Conforms Solubility Insoluble in water, Conforms miscible with oilsFatty Acid Profile [%] VQP-050 (GC) Palmitic Acid 6.3 Stearic Acid 1.9Oleic Acid 22.3 Linoleic Add 9.7 α-Linolenic Acid >56 59.8 Total FattyAcids (% w/w) 85 . . . 95 88.6 Peroxide Value (meqO₂/kg) <10 VQP-049 2.3Water Content (%)   <1.5 VQP-048 <0.2 Heavy Metals (ppm) <10 ICP-MS <1Microbiological Data (cfu/g) Total Aerobic Microbial Count  <10³ USP<61> <100 Combined Yeast & Mold  <10² USP <61> <100 E. coli/TotalColiforms <10 AOAC 991.14 <10 {all values as is basis} All of theingredients are GMO free. Therefore this product is in accordance withEU regulations 1830/2003 and 1829/2003. The Product has not been treatedwith gamma rays.

Table 2 show accelerated stability testing of the perilla seed extractwith an OTB® per oxidation blocker and Table 3 shows the acceleratedstability testing of a perilla seed extract sample produced by ValensaInternational of Eustis, Fla. as compared to chia seed extracts such asthe Chia Gold™ which is produced by the process shown in FIG. 1.

TABLE 2 Accelerated Stability Testing of Valensa Perilla Seed Extractwith O2B ® Rancidity Induction Time Shelf Life @20° C. (hr)¹ (yr)²Perilla Oil (without O2B) 7 0.5 Perilla Oil (with O2B) 42 2.8 ¹Rancimatinduction time measured in hours with air bubbling through heated oil(90° C.) in the light. ²Rancimat data is a function of accelerated highheat and oxygen exposure. Extrapolated data plots are assumed linear inthe presence of air and light at 20° C. however, if product is stored at20° C. in an air and light barrier package, then shelf stability is atleast doubled.

TABLE 3 Accelerated Stability Testing of Valensa Perilla Seed and ChiaSeed Extracts with O2B ® Rancidity Shelf Life Shelf Life Induction Time@20° C. Increase (hr)¹ (yr)² (%) Perilla Oil 7 0.5 (without O2B) PerillaOil 42 2.8 611% (with O2B) Tresalbio Chia Oil 12 0.4 (without O2B)Tresalbio Chia Oil 73 2.2 603% (with O2B) ChiaGold Oil 9 0.3 (withoutO2B) ChiaGold Oil 60 1.8 663% (with O2B) ¹Rancimat induction timemeasured in hours with air bubbling through heated oil (80-90° C.) inthe light. ²Rancimat data is a function of accelerated high heat andoxygen exposure. Extrapolated data plots are assumed linear in thepresence of air and light at 20° C. however, if product is stored at 20°C. in an air and light barrier package, then shelf stability is at leastdoubled.

Table 1 illustrates various values and shows the total fatty acids (%w/w) is 85 to about 95 and has in that particular example a result of88.6. It should be understood that the seed oil could possibly have aslow as about 60% w/w of PUFAs and as high as about 95% and a ratio offrom about 4:1 to about 6:1 ALA to LA. The peroxide value of the seedoil is typically under 10.0 meq/Km. The PUFAs typically comprise atleast greater than 50% ALA and in the example shown in Table 1 isgreater than 56% and in one particular example shown in FIG. 1 is 59.8.The seed oil is shelf stable at room temperature up to 32 months in aparticular example. Other data is shown such as specific components ofthe fatty acids and the water content, heavy metals in PPM, andmicrobiological data in CFU/G, such as the total aerobic microbialcount, a combined yeast and mold and E. coli/total coliforms. All thevalues are an as-is basis and the ingredients are GMO free. Therefore,this product is in accordance with EU regulations 1830/2003 and1829/2003. This product had not been treated with gamma rays.Ingredients include the perilla (perilla frutescens (L) Britton) seedoil, OTB® botanical per oxidation blocker including refined non-GMOsoybean mixed tocopherols and spice extracts. GMO corresponds togenetically modified organisms, and thus, non-GMO refers tonon-genetically modified organisms. The soybean had not been createdthrough gene-splicing techniques of biotechnology or geneticengineering.

The production diagram in FIG. 2 shows the process used to obtain theperilla seed extract in accordance with a non-limiting example. Theextraction technology has been described relative to the chia seed oiland extract in the incorporated by reference parent application anddescribed above in some detail. It should be understood that extractiontechnology creates materials for human nutrition and supplementation andoffers various benefits including enhanced efficacy with the isolationof key components to allow higher dosage and targeted performance.Extraction allows standardization. Natural materials tend to vary inmake-up and extraction makes them consistent. It is also convenientbecause smaller dosages of high efficiency materials allow a consumer tomore easily obtain the required levels of nutrients in a daily regimen.There is also enhanced safety because extraction gives more of thedesired products and less of what is not desired. Extraction allows theremoval of compounds that are not optimal for human health from naturalmaterials.

The desired extraction technology uses the DEEP EXTRACT® process fromValensa International as an ultra high pressure carbon dioxideextraction technology that yields micronutrients and has high extractionefficiency to deliver more of the higher molecular weight compounds thatmore closely track the natural source materials. The process is flexibleand allows for possible fractionation of the product if desired anddelivery of specific compounds out of the raw material.

This extraction process, such as the DEEP EXTRACT@ process, offers amore gentle treatment of high value raw materials at temperature levelsthat are substantially below those used in other expeller pressprocesses and some chemical solvent extraction processes in the absenceof oxygen. This reduces the degradation of liable compounds, chemicalchange of a component and the oxidation potential. The supercritical CO²process offers virtual sterilization of the finished product andbiomass, which are untouched by chemical solvents and stay “natural” asbefore extraction. Selective fractionation in some example isadvantageous. Pressure is a main tool used to tailor the resultingfractionated products for product quality and efficient manufacturing.Because the CO² extraction is an all-natural and organic process, thefinal product is devoid of impurities and residues and delivers throughthe supercritical CO² extraction the high molecular weight compoundssuch as sterols, carotenoids and long chain alcohols.

Supercritical CO² extraction is advantageous over other methods used forextracting botanicals, including tinctures (usually alcohol extraction;steam distillation; expeller pressing, sometimes referred to as “coldpressing,” and chemical solvent extraction). Chemical solvent extractiontechnology using strong solvents and supercritical CO² technology usinghigh pressures typically offer the most comprehension extraction of abotanical. Supercritical CO² extraction conducted under very highpressure is advantageous. When carbon dioxide gas (CO²) is compressedabove 73 bar at a temperature above 31 degrees C. (87.8 degreesFahrenheit), it transforms into a dense gas as supercritical CO², whichhas an extremely high solvating capacity and a power to extractconstituents of botanicals. Its solvating capacity is a function of itsdensity and by changing its density with pressure, the manufacturer isable to select the quality, quantity and specific principles of thetargeted extract. Supercritical CO² is biologically compatible andgenerally regarded as safe (GRAS) by the FDA. It is also non-flammableand environmentally sound.

Any defatted cake resulting from the supercritical CO² process istypically viable and can be marketed or used for further processing in awide range of human/food applications. Sometimes the defatted cake isthe primary product and the oil is secondary as described in commonlyassigned U.S. patent application Ser. No. 12/349,100 as filed on Jan. 6,2009 and published as U.S. Patent Publication No. 2009/0181127, thedisclosure which is hereby incorporated by reference in its entirety.That application describes a chia seed composition as a composition ofmatter formed from a stable, defatted whole grain flour derived fromsalvia hispanica L. whole ground seed using a suitable solvent such asusing supercritical CO² fluid extraction. This chia seed extractiondemonstrates that after processing of supercritical CO², the extractcake contains in some examples virtually no fat or oil and the resultantpowder is approximately 50% protein and 50% carbohydrates that existessentially as fiber.

The supercritical CO² process offers a gentle treatment of high valueraw materials at temperature levels substantially below those used inexpeller press in some chemical solvent extraction operations in theabsence of oxygen. This reduces the degradation of labile compounds,chemical changing components and the oxidation potential. Thesupercritical CO² process offers virtual sterilization of the finishedproduct and biomass that are untouched by chemical solvents and stay“natural” as they were before extraction. It also allows the advantageof fractionating the extracted components selectively with pressuretailoring the resulting fractionated products for product quality andefficient manufacturing. This addresses pesticide/insecticide residuesand handle concerns about microorganisms that are present in expellerpress materials.

The extract as shown in FIG. 2 is dewatered and decantated andantioxidants added such as the Valensa OTB® per oxidation blocker systemas a stabilizer to ensure that the botanical extract reaches a consumerin an efficacious and safe form. Stabilization with the OTB® componentsis a key to shelf life and continued product quality and is advantageousover using preservatives to stabilize natural materials, which is oftenseen as a negative by consumers. The OTB® per oxidation blocker systemused by Valensa is 100% natural, non-GMO, and protects sensitive oilsand particularly the highly unsaturated oils derived from fish andbotanicals from the manufacture to consumption. The OTB® per oxidationblocker is a synergistic proprietary formulation of powerful naturalcompounds including astaxanthin, phenolic antioxidants and naturaltocopherols. This technology prevents destructive oxidative,photochemical and rancification reactions. It protects expensive andsensitive compounds such as carotenoids and polyunsaturated fatty acidsand can boost the effectiveness of other antioxidants such as vitamin Ebecause it chemically quenches stable vitamin E free radicals. Theantioxidants have in-vivo activity to protect both products and people.

Perilla has a number of essential oils. These are extracted from theleaves of perilla. About 50% to about 60% of perillaldehyde isresponsible for so much of the aroma and taste of perilla. There areother terpenes such as limonene, caryophyllene and farnesene. There areother chemotypes such as perilla keytone (PK), esholzia keytone (EK),perillene (PL), and various phenylpropanoids such as myristicin,dillapiole and elemicin. Citral is a type rich in rosefuran. Perilla oiltypically is obtained by pressing the seeds of perilla that containabout 35% to about 45% oil. In some parts of Asia, perilla oil is anedible oil valued for medicinal benefit. Typically, perilla oil is arich source of omega-3 fatty acid alpha-linolenic acids. As a dryingoil, it is similar to tung oil or linseed oil and is sometimes used inpaint, varnish, linoleum, printing ink, lacquers and other protectivewaterproof coatings. In Japan, the oxime of perillaldehyde (perillartin)is used as an artificial sweetener and typically is about 2,000 timessweeter than sucrose.

Rancimat testing has shown the advantages of the perilla seed oilextract in accordance with a non-limiting example. This method is anaccelerated oxidation test that is a simple, quick and efficient way toscreen the effectiveness of the antioxidants used in liquid fats andoils. Typically, the rancimat test is an accelerated oxidation test inwhich the oil or fat to be tested is run at an elevated temperatureexposing the sample to air accelerating the oxidation process of theoil. Auto oxidation typically occurs in a few hours instead of themonths or years and the metabolites are driven off into a measuringvessel that measures the change in conductivity in one example. Thiswould indicate the point at which the formation of volatile carboxylicacids and oxidation has occurred.

It is also possible to disperse nano- and/or micro-particles of rice orsugar cane based policosanol for providing a heart healthy dietarysupplement. Such dietary supplement composition additives are disclosedin commonly assigned U.S. Pat. No. 7,959,950, the disclosure which ishereby incorporated by reference in its entirety. This human or animaldietary supplement composition includes one or more long chain (C24-C36)primary alcohols (policosanols) dispersed in food-grade oils or fatswhere the average policosanol particle size is greater than 2 micronsand less than 10 microns in one example, and in another example, lessthan 100 microns.

Many modifications and other embodiments of the invention will come tothe mind of one skilled in the art having the benefit of the teachingspresented in the foregoing descriptions and the associated drawings.Therefore, it is to be understood that the invention is not to belimited to the specific embodiments disclosed, and that themodifications and embodiments are intended to be included within thescope of the dependent claims.

1-25. (canceled)
 26. A method of manufacturing a perilla frutescensderived seed oil comprising from 60 to 95 percent w/w of PUFAs in aratio of from 4:1 to 6:1 alpha-linolenic acid (ALA) to linoleic acid(LA), comprising: processing perilla frutescens seed in the absence ofoxygen to obtain a biomass having a desired particle size distribution;subjecting the resulting biomass to supercritical fluid CO² extraction;collecting any resulting seed oil fractions; and separating water ineach fraction.
 27. The method according to claim 26, and furthercomprising treating any resulting seed oil fractions with antioxidantsto impart a desired room temperature stability.
 28. The method accordingto claim 26, and further comprising controlling the extent of oilextraction by particle size distribution.
 29. The method according toclaim 26, and further comprising adding propane in admixture withsupercritical CO² in the supercritical state as an extraction solvent.30. The method according to claim 26, and further comprising addinglipophilic antioxidants to increase the room temperature stability ofthe resulting oil.
 31. The method according to claim 26, and furthercomprising adding lipophilic antioxidants either alone or in combinationwith at least one of: a) phenolic antioxidants including at least one ofsage, oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d)carotenoids including at least one of astaxanthin, lutein, andzeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g) Butylatedhydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i) TertiaryButyl hydroquinone (TBHQ).
 32. The method according to claim 26, andfurther comprising treating any resulting dewatered seed oil withbleaching clay or activated carbon.
 33. The method according claim 26,and further comprising adding a hydrophilic antioxidant or sequesterantcomprising hydrophilic phenolic antioxidants including at least one ofgrape seed extract, tea extracts, ascorbic acid, citric acid, tartaricacid, and malic acid.
 34. The method according to claim 26, and furthercomprising premilling or roller press flaking the perilla frutescensseed to obtain the desired particle size.
 35. The method according toclaim 34, and further comprising treating the premilled or roller pressflaked seed with a lipophilic or hydrophilic antioxidant prior tosolvent extraction.
 36. The method according to claim 26, and furthercomprising processing the perilla frutescens seed with or without theaddition of hydrophilic or lipophilic antioxidants during the particlesizing process.
 37. The method according to claim 26, wherein saidperilla frutescens derived seed oil is stable at room temperature for upto 32 months.
 38. The method according to claim 26, and furthercomprising performing supercritical extraction in the presence oflipophilic and or hydrophilic antioxidants.
 39. A method ofmanufacturing a perilla frutescens derived seed oil comprising from 60to 95 percent w/w of PUFAs in a ratio of from 4:1 to 6:1 alpha-linolenicacid (ALA) to linoleic acid (LA) that is stable at room temperature forup to 32 months, comprising: pre-milling or roller press flaking perillafrutescens seed in the absence of oxygen to obtain a cracked seedbiomass having a desired particle size distribution; extracting seed oilfrom the cracked seed biomass by subjecting the cracked seed biomass tosupercritical fluid CO² extraction; fractionating the resulting seed oilextract and collecting light and heavy fractions of the seed oilextract; and separating any remaining water in at least one of the lightand heavy fractions of seed oil extract.
 40. The method according toclaim 39, and further comprising treating light and heavy fractions ofseed oil extract with antioxidants to impart a desired room temperaturestability.
 41. The method according to claim 39, and further comprisingcontrolling the extent of oil extraction by pre-milling or roller pressflaking the seed to a desired particle size.
 42. The method according toclaim 39, and further comprising adding propane in admixture withsupercritical CO² in the supercritical state as an extraction solvent.43. The method according to claim 39, and further comprising addinglipophilic antioxidants to increase the room temperature stability ofthe resulting oil.
 44. The method according to claim 39, and furthercomprising adding lipophilic antioxidants either alone or in combinationwith at least one of: a) phenolic antioxidants including at least one ofsage, oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d)carotenoids including at least one of astaxanthin, lutein, andzeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g) Butylatedhydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i) TertiaryButyl hydroquinone (TBHQ).
 45. The method according to claim 39, andfurther comprising treating any light and heavy fractions of seed oilextract with bleaching clay or activated carbon.
 46. The methodaccording claim 39, and further comprising adding a hydrophilicantioxidant or sequesterant comprising hydrophilic phenolic antioxidantsincluding at least one of grape seed extract, tea extracts, ascorbicacid, citric acid, tartaric acid, and malic acid.
 47. The methodaccording to claim 39, and further comprising treating the pre-milled orroller press flaked seed with a lipophilic or hydrophilic antioxidantprior to solvent extraction.
 48. The method according to claim 39, andfurther comprising performing supercritical extraction in the presenceof lipophilic and/or hydrophilic antioxidants.